INSERM, RESINFIT, U1092, Limoges, France.
Université Limoges, RESINFIT, U1092, Limoges, France.
PLoS Genet. 2022 Apr 28;18(4):e1010177. doi: 10.1371/journal.pgen.1010177. eCollection 2022 Apr.
Class 1 integrons are widespread genetic elements playing a major role in the dissemination of antibiotic resistance. They allow bacteria to capture, express and exchange antibiotic resistance genes embedded within gene cassettes. Acquisition of gene cassettes is catalysed by the class 1 integron integrase, a site-specific recombinase playing a key role in the integron system. In in vitro planktonic culture, expression of intI1 is controlled by the SOS response, a regulatory network which mediates the repair of DNA damage caused by a wide range of bacterial stress, including antibiotics. However, in vitro experimental conditions are far from the real lifestyle of bacteria in natural environments such as the intestinal tract which is known to be a reservoir of integrons. In this study, we developed an in vivo model of intestinal colonization in gnotobiotic mice and used a recombination assay and quantitative real-time PCR, to investigate the induction of the SOS response and expression and activity of the class 1 integron integrase, IntI1. We found that the basal activity of IntI1 was higher in vivo than in vitro. In addition, we demonstrated that administration of a subinhibitory concentration of ciprofloxacin rapidly induced both the SOS response and intI1 expression that was correlated with an increase of the activity of IntI1. Our findings show that the gut is an environment in which the class 1 integron integrase is induced and active, and they highlight the potential role of integrons in the acquisition and/or expression of resistance genes in the gut, particularly during antibiotic therapy.
1 类整合子是广泛存在的遗传元件,在抗生素耐药性的传播中起着重要作用。它们允许细菌捕获、表达和交换嵌入基因盒中的抗生素耐药基因。基因盒的获取由 1 类整合子整合酶催化,整合酶是一种位点特异性重组酶,在整合子系统中起着关键作用。在体外浮游培养中,intI1 的表达受 SOS 反应调控,SOS 反应是一个调节网络,介导由多种细菌应激引起的 DNA 损伤修复,包括抗生素。然而,体外实验条件与肠道等自然环境中细菌的真实生活方式相去甚远,肠道是整合子的储存库。在本研究中,我们在无菌小鼠中建立了肠道定植的体内模型,并使用重组测定和实时定量 PCR 来研究 SOS 反应的诱导以及 1 类整合子整合酶 IntI1 的表达和活性。我们发现,IntI1 的基础活性在体内高于体外。此外,我们证明,给予亚抑菌浓度的环丙沙星可迅速诱导 SOS 反应和 intI1 的表达,这与 IntI1 活性的增加相关。我们的研究结果表明,肠道是整合酶被诱导和活跃的环境,并强调了整合子在肠道中获取和/或表达耐药基因的潜在作用,特别是在抗生素治疗期间。